LOCUS       sequence-113790-        7479 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast expression plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7479)
  AUTHORS   Chen Y, Daviet L, Schalk M, Siewers V, Nielsen J
  TITLE     Establishing a platform cell factory through engineering of yeast 
            acetyl-CoA metabolism.
  JOURNAL   Metab Eng. 2013 Jan;15:48-54. doi: 10.1016/j.ymben.2012.11.002. Epub
            2012 Nov 17.
  PUBMED    23164578
REFERENCE   2  (bases 1 to 7479)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Metab 
            Eng."; date: "2013-01"; volume: "15:48-54. doi"; pages: " 
            10.1016/j.ymben.2012.11.002. Epub 2012 Nov 17"
COMMENT     Sequence Label: pIYC04
FEATURES             Location/Qualifiers
     source          1..7479
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        317..503
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             504..1163
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALLARGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHFL
                     ESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     rep_origin      complement(1423..1878)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1560..1581)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1772..1791
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(1987..2152)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     2073..2097
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(2300..2323)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(2357..2763)
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     3765..3784
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3765..3783
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3801..3830
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     terminator      3860..4049
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4148..4165)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4319..4907)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4399..4418)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5078..5869,5870..5938))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5701..5720
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5939..6043)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6070..7412
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(7033..7080)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     7452..7470
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataaatt cccgttttaa gagcttggtg agcgctagga gtcactgcca ggtatcgttt
      241 gaacacggca ttagtcaggg aagtcataac acagtccttt cccgcaattt tctttttcta
      301 ttactcttgg cctcctctag tacactctat atttttttat gcctcggtaa tgattttcat
      361 tttttttttt cccctagcgg atgactcttt ttttttctta gcgattggca ttatcacata
      421 atgaattata cattatataa agtaatgtga tttcttcgaa gaatatacta aaaaatgagc
      481 aggcaagata aacgaaggca aagatgacag agcagaaagc cctagtaaag cgtattacaa
      541 atgaaaccaa gattcagatt gcgatctctt taaagggtgg tcccctagcg atagagcact
      601 cgatcttccc agaaaaagag gcagaagcag tagcagaaca ggccacacaa tcgcaagtga
      661 ttaacgtcca cacaggtata gggtttctgg accatatgat acatgctctg gccaagcatt
      721 ccggctggtc gctaatcgtt gagtgcattg gtgacttaca catagacgac catcacacca
      781 ctgaagactg cgggattgct ctcggtcaag cttttaaaga ggccctactg gcgcgtggag
      841 taaaaaggtt tggatcagga tttgcgcctt tggatgaggc actttccaga gcggtggtag
      901 atctttcgaa caggccgtac gcagttgtcg aacttggttt gcaaagggag aaagtaggag
      961 atctctcttg cgagatgatc ccgcattttc ttgaaagctt tgcagaggct agcagaatta
     1021 ccctccacgt tgattgtctg cgaggcaaga atgatcatca ccgtagtgag agtgcgttca
     1081 aggctcttgc ggttgccata agagaagcca cctcgcccaa tggtaccaac gatgttccct
     1141 ccaccaaagg tgttcttatg tagtgacacc gattatttaa agctgcagca tacgatatat
     1201 atacatgtgt atatatgtat acctatgaat gtcagtaagt atgtatacga acagtatgat
     1261 actgaagatg acaaggtaat gcatcattct atacgtgtca ttctgaacga ggcgcgcttt
     1321 ccttttttct ttttgctttt tctttttttt tctcttgaac tcgacggatc tatgcggtgt
     1381 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggaaattgta aacgttaata
     1441 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     1501 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     1561 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     1621 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     1681 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     1741 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     1801 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     1861 cgccgctaca gggcgcgtcg cgccattcgc cattcaggct gcgcaactgt tgggaagggc
     1921 gatcggtgcg ggcctcttcg ctattacgcc agctggataa aggcgcgcca aacgacctag
     1981 gaattggagc gacctcatgc tatacctgag aaagcaacct gacctacagg aaagagttac
     2041 tcaagaataa gaattttcgt tttaaaacct aagagtcact ttaaaatttg tatacactta
     2101 ttttttttat aacttattta ataataaaaa tcataaatca taagaaattc gcttatttag
     2161 aagtgtcaac aacgtatcta ccaacgattt gacccttttc catcttttcg taaatttctg
     2221 gcaaggtaga caagccgaca accttgattg gagacttgac caaacctctg gcgaagaatt
     2281 gttaattaag agctcagatc ttatcgtcgt catccttgta atccatcgat actagtgcgg
     2341 ccgcttgtaa ttaaaactta gattagattg ctatgctttc tttctaatga gcaagaagta
     2401 aaaaaagttg taatagaaca agaaaaatga aactgaaact tgagaaattg aagaccgttt
     2461 attaacttaa atatcaatgg gaggtcatcg aaagagaaaa aaatcaaaaa aaaaaatttt
     2521 caagaaaaag aaacgtgata aaaattttta ttgccttttt cgacgaagaa aaagaaacga
     2581 ggcggtctct tttttctttt ccaaaccttt agtacgggta attaacgaca ccctagagga
     2641 agaaagaggg gaaatttagt atgctgtgct tgggtgtttt gaagtggtac ggcgatgcgc
     2701 ggagtccgag aaaatctgga agagtaaaaa aggagtagaa acattttgaa gctatggtgt
     2761 gtgcggccgg cctggaagta ccttcaaaga atggggtctt atcttgtttt gcaagtacca
     2821 ctgagcagga taataataga aatgataata tactatagta gagataacgt cgatgacttc
     2881 ccatactgta attgctttta gttgtgtatt tttagtgtgc aagtttctgt aaatcgatta
     2941 attttttttt ctttcctctt tttattaacc ttaattttta ttttagattc ctgacttcaa
     3001 ctcaagacgc acagatatta taacatctgc ataataggca tttgcaagaa ttactcgtga
     3061 gtaaggaaag agtgaggaac tatcgcatac ctgcatttaa agatgccgat ttgggcgcga
     3121 atcctttatt ttggcttcac cctcatacta ttatcagggc cagaaaaagg aagtgtttcc
     3181 ctccttcttg aattgatgtt accctcataa agcacgtggc ctcttatcga gaaagaaatt
     3241 accgtcgctc gtgatttgtt tgcaaaaaga acaaaactga aaaaacccag acacgctcga
     3301 cttcctgtct tcctattgat tgcagcttcc aatttcgtca cacaacaagg tcctagcgac
     3361 ggctcacagg ttttgtaaca agcaatcgaa ggttctggaa tggcgggaaa gggtttagta
     3421 ccacatgcta tgatgcccac tgtgatctcc agagcaaagt tcgttcgatc gtactgttac
     3481 tctctctctt tcaaacagaa ttgtccgaat cgtgtgacaa caacagcctg ttctcacaca
     3541 ctcttttctt ctaaccaagg gggtggttta gtttagtaga acctcgtgaa acttacattt
     3601 acatatatat aaacttgcat aaattggtca atgcaagaaa tacatatttg gtcttttcta
     3661 attcgtagtt tttcaagttc ttagatgctt tctttttctc ttttttacag atcatcaagg
     3721 aagtaattat ctacttttta caacaaatat aaaacaagga tccgtaatac gactcactat
     3781 agggcccggg cgtcgacatg gaacagaagt tgatttccga agaagacctc gagtaagctt
     3841 ggtaccgcgg ctagctaaga tccgctctaa ccgaaaagga aggagttaga caacctgaag
     3901 tctaggtccc tatttatttt tttatagtta tgttagtatt aagaacgtta tttatatttc
     3961 aaatttttct tttttttctg tacagacgcg tgtacgcatg taacattata ctgaaaacct
     4021 tgcttgagaa ggttttggga cgctcgaaga tcctccggat cgtttcgccg gcgtttatcc
     4081 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     4141 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     4201 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     4261 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     4321 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     4381 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     4441 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     4501 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     4561 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     4621 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     4681 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     4741 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     4801 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     4861 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     4921 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     4981 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     5041 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     5101 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     5161 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     5221 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     5281 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     5341 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     5401 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     5461 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     5521 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     5581 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     5641 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     5701 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     5761 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     5821 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     5881 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     5941 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     6001 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     6061 tgccacctga acgaagcatc tgtgcttcat tttgtagaac aaaaatgcaa cgcgagagcg
     6121 ctaatttttc aaacaaagaa tctgagctgc atttttacag aacagaaatg caacgcgaaa
     6181 gcgctatttt accaacgaag aatctgtgct tcatttttgt aaaacaaaaa tgcaacgcga
     6241 gagcgctaat ttttcaaaca aagaatctga gctgcatttt tacagaacag aaatgcaacg
     6301 cgagagcgct attttaccaa caaagaatct atacttcttt tttgttctac aaaaatgcat
     6361 cccgagagcg ctatttttct aacaaagcat cttagattac tttttttctc ctttgtgcgc
     6421 tctataatgc agtctcttga taactttttg cactgtaggt ccgttaaggt tagaagaagg
     6481 ctactttggt gtctattttc tcttccataa aaaaagcctg actccacttc ccgcgtttac
     6541 tgattactag cgaagctgcg ggtgcatttt ttcaagataa aggcatcccc gattatattc
     6601 tataccgatg tggattgcgc atactttgtg aacagaaagt gatagcgttg atgattcttc
     6661 attggtcaga aaattatgaa cggtttcttc tattttgtct ctatatacta cgtataggaa
     6721 atgtttacat tttcgtattg ttttcgattc actctatgaa tagttcttac tacaattttt
     6781 ttgtctaaag agtaatacta gagataaaca taaaaaatgt agaggtcgag tttagatgca
     6841 agttcaagga gcgaaaggtg gatgggtagg ttatataggg atatagcaca gagatatata
     6901 gcaaagagat acttttgagc aatgtttgtg gaagcggtat tcgcaatatt ttagtagctc
     6961 gttacagtcc ggtgcgtttt tggttttttg aaagtgcgtc ttcagagcgc ttttggtttt
     7021 caaaagcgct ctgaagttcc tatactttct agagaatagg aacttcggaa taggaacttc
     7081 aaagcgtttc cgaaaacgag cgcttccgaa aatgcaacgc gagctgcgca catacagctc
     7141 actgttcacg tcgcacctat atctgcgtgt tgcctgtata tatatataca tgagaagaac
     7201 ggcatagtgc gtgtttatgc ttaaatgcgt acttatatgc gtctatttat gtaggatgaa
     7261 aggtagtcta gtacctcctg tgatattatc ccattccatg cggggtatcg tatgcttcct
     7321 tcagcactac cctttagctg ttctatatgc tgccactcct caattggatt agtctcatcc
     7381 ttcaatgcta tcatttcctt tgatattgga tcatctaaga aaccattatt atcatgacat
     7441 taacctataa aaataggcgt atcacgaggc cctttcgtc
//
